Collecting Plasma From Whole Blood

Abstract


The protocol is derived from the WHO/IARC guideline “Common Minimal Technical Standards and Protocols”. The purpose of the document is to provide a framework for the development and coordination of Biological Resource Centers.
Plasma collected in EDTA and ACD tubes, can be used for bioassays, plasma DNA isolation, proteomic analysis, and biomarker discovery.
If serum and plasma are being collected, it is important to prioritize the separation of them so they can be frozen as soon as possible. This is critical for time sensitive samples for protein studies for example.

Equipment

  • EtOH , to clean tubes
  • sterile 15ml centrifuge tubes , nr depending on amount of blood
  • Cryovial , for snap freezing
  • Liquid Nitrogen , for snap freezing and possibly storage
  • Freezer (-80) , or liquid nitrogen storage
  • Centrifuge , compliant with 15ml centrifuge tubes

Reagents

  • See equipment

Instructions

  1. Spin vacutainer (about 9ml) at 815xg for 10 minutes at 4°C to separate plasma from blood cells.
  2. After wiping each tube with 70% alcohol, remove about 3ml plasma.
  3. Tube can be retained for white blood cell extraction.
  4. Transfer to a labeled 15ml tube and centrifuge at 2500xg for 10 minutes at 4°C.
  5. The purpose of double spinning the plasma is to remove all cellular contaminants so that the plasma is suitable for plasma DNA analysis. It is extremely important, therefore, not to disturb the buffy coat after the first spin, and any pellet after the second spin.
  6. Aliquot plasma into 1ml labeled cryovials (3 to 4 aliquots).
  7. Place in liquid nitrogen Dewar to snap freeze.
  8. Transfer to a labeled 15ml tube and centrifuge at 2500xg for 10 minutes at 4°C.
  9. Store at -80°C or in liquid nitrogen.

Citation:
IARC -International agency for research on cancer. Collecting Plasma From Whole Blood. The Molecular Methods database. Wed, 12/19/2012 - 11:26. Acc.nr AZgLt10.